Inhibition of Ceramide Metabolism Sensitizes Human Leukemia Cells

Background Adar2 deaminates selective adenosines to inosines (A-to-I RNA editing) in the double-stranded region of nuclear transcripts. neuronal populations nor rescued the developmental defects. The expressions of and in the neural crest cells were reduced in the and restored normal expressions of and and expressions. These results argue that an elevated GluA2Q level is sufficient for generating the cranial neural crest defects observed in the (is usually fully edited at the Q/R site throughout mouse development. The edited R form (GluA2R) subunit plays a dominant role in reducing the MCDR2 Ca2+ entry of GluA2R-containing AMPARs [9]. Mice with a Q/R editing-deficient allele of (mouse and the abnormalities are rescued by replacement of the chromosomal with at the Q/R site is responsible for the abnormalities of lacking the homolog displays age-dependent neurological and behavior defects but is usually morphologically normal with normal lifespan under optimal conditions [11]. Mice defective in are embryonic lethal, display defective hematopoiesis and widespread apoptosis in tissues expressing high levels of have been identified [13], [14]. A-to-I editing of zebrafish and kainate Hyodeoxycholic acid receptor subunit has also been reported [15]C[17]. Interestingly, the editing of during early zebrafish development is usually incomplete [16] and the chromosomal sequence of the other paralogue, paralogues of more derived teleost carry chromosomally encoded R codon [15]. In this study, we demonstrate an evolutionarily conserved function of zebrafish Adar2 in editing the Q/R site of Reducing expression and reducing Q/R editing of resulted in severe developmental defects in the nervous system and cranial cartilages. Further studies revealed that the induction of apoptosis and reduced number of spinal cord motor neurons in the morphants depended on p53, while the developmental defects in brain, lateral line neuromasts and head cartilages were p53-impartial. Results of overexpressing the edited and unedited forms of GluA2 in the morphant and wild type zebrafish embryos demonstrate that an elevation of the unedited GluA2Q level is sufficient to disturb the development of neural crest cells in zebrafish. Results Expression pattern of transcript in the 1-cell (0 hpf) and blastrula-staged (4 hpf) embryos, indicating that maternal transcript was presented in the zebrafish embryos. The level (relative to the level of transcript decreased at 10 hpf and then remained stable between 10 to 72 hpf (Fig. S1). WISH (whole-mount hybridization) analysis revealed that was ubiquitously expressed in the epiblast during gastrulation and early segmentation periods. Slightly higher expressions of were detected in the neural plate of Hyodeoxycholic acid bud-stage embryos (Fig. 1A and D) as well as in the hindbrain (hb) and somites of 6-somite stage embryos (Fig. 1B and E). The expression of became more restricted to the nervous system at later segmentation stages (Figs. 1C and F). Persistent expression of in the forebrain (telecephalon and diencephalon), retina and cranial Hyodeoxycholic acid sensory ganglia was maintained between 24 to 72 hpf (Figs. 1G-P), while expression of in the caudal region of CNS (hindbrain and spinal cord) decreased after 36 hpf. The expression of in the ventral midbrain (tegmentum) became more prominent at 30 hpf (Fig. 1I). At 48-hpf, enriched expression of was observed in discrete areas of ventral midbrain, matching the locations of cranial motor Hyodeoxycholic acid neurons (asterisks, Fig. 1N). In addition to the expression in the nervous system, was highly expressed in the heart (Figs. 1K, M, O, and P) and the third to seventh pharyngeal arches (cb 1C5, Fig. 1O and P). Low levels of expression were also detected in the fin bud/pectoral Hyodeoxycholic acid fin, liver and digestive tract (Fig. 1L, N, P and P). Physique 1 Expression patterns of zebrafish during embryogenesis. In general, the expression domains of in the CNS and cranial sensory neurons overlapped with that of the AMPAR subunit genes, and and a putative substrate of Adar2, were not identical. By quantitative RT-PCR analysis, the expression of has been reported to significantly increase after 30 hpf [16], while that of mildly decrease (Fig. S1). Robust expression of in the retina and cranial ganglia, especially the posterior lateral line ganglion/placode, started at 24 hpf (Fig. 1G), earlier than an overt expression of in these regions [18]. After 36 hpf, the expression in the spinal cord and medulla oblongata diminished, while expression persisted (Fig. 1K and L). Moreover, expression has not been reported in the pharyngeal cartilages. Reduction of Q/R RNA editing of in and was estimated by RT-PCR (Fig. 2B). The splicing of was not affected before midblastrula transition (4 hpf) when.